Composition for hardening soft tissue

ABSTRACT

The present invention relates to various compounds capable of temporarily hardening soft tissue for surgical suturing of the soft tissue. The compounds according to the present invention can temporarily increase the hardness or tension of soft tissue, thereby improving the suturing efficiency during suturing of the soft tissue, thereby preventing aftereffects or the like from occurring due to insufficient anastomosis. In addition, the compounds according to the present invention can temporarily increase the hardness or tension of soft tissue, particularly pancreas, thereby increasing the suturing efficiency during pancreaticoduodenectomy and effectively preventing pancreatic leakage.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. Patent ApplicationSer. No. 16/325,073, filed Feb. 12, 2019, which is a national stageapplication under 35 U.S.C. 371 and claims the benefit of PCTApplication No. PCT/KR2017/008743 having an international filing date of11 Aug. 2017, which designated the United States, which PCT applicationclaimed the benefit of Republic of Korea Application No.10-2016-0102895, filed 12 Aug. 2016, the disclosures of each of whichare incorporated herein by reference in their entireties.

TECHNICAL FIELD

The present invention relates to compounds capable of temporarilyhardening soft tissue for surgical suturing of the soft tissue.

BACKGROUND ART

Pancreas is an organ located behind the stomach. It secretes a varietyof digestive enzymes which promote degradation of the carbohydrates,proteins and fats that passed through the stomach. Also, it producesimportant hormones such as insulin, glucagon and the like. If thesecretion of digestive enzymes from the pancreas is insufficient, theuptake of nutrients becomes insufficient. In addition, the secretion ofinsulin and glucagon regulates intravascular glucose levels. Studies onthe mechanisms that control the production and secretion of thesevarious enzymes and hormones have become important topics in relation tonutrition and diverse diseases, such as diabetes, obesity and the like.

Major diseases that occur in relation to the pancreas include pancreaticcancer, pancreatitis, diabetes and the like. Diabetes is a risky diseasethat occurs when insulin secretion in the pancreas declines orresistance to insulin action in other tissues occurs, leading to variouscomplications due to elevated blood glucose levels. In addition,pancreatic cancer is known to be a disease that has the highestmortality rate among malignant tumors and has a poor prognosis.

The most common treatment methods for the treatment of diseases relatedto the pancreas are performed by the administration or intravenousinfusion of analgesics, antibiotics, anticancer drugs, or the like. Insevere cases, pancreaticoduodenectomy is performed, which treats thediseases by surgery.

The pancreaticoduodenectomy has been widely distributed by Whipple etal. since successful pancreaticoduodenectomy was first introduced byKausch in 1912. The pancreaticoduodenectomy is an important surgicalprocedure that is applied to radical treatment of Vater periampullarycancer which occurs in the pancreas, head, common bile duct, duodenalampulla or the like, but it is a surgical method having a highpossibility of developing postoperative complications. Until the 1970s,the pancreaticoduodenectomy was reported to have a postoperativemortality rate of about 20%. Recently, pancreaticoduodenectomy has beenreported to have a mortality rate of 5% or less due to advances insurgical technology, anesthetic technology, patient management beforeand after surgery, nutritional management, and the like.

However, despite the dramatic decrease in mortality rate, the incidenceof postoperative complications has been reported to be 40-50% even inrecent years. Among these complications, the incidence of pancreaticleakage, which is closely related to mortality rate, has been reportedto be still 10-20%. It was found that pancreatic leakage, when occurred,could cause intraperitoneal hemorrhage, abscess or the resulting sepsis,and the mortality rate associated with it was also high.

According to several reports, it is known that the diameter of thepancreatic duct, the consistency of the pancreatic parenchyma, and thelike, are risk factors that are involved in the occurrence of pancreaticleakage. Here, regarding the consistency of the pancreatic parenchyma,the pancreas parenchyma remaining after pancreatectomy is very soft, sothere is a high risk of crushing the tissue at the time of suturing, andhence a high level of skill is required for suturing. Even after thesuturing, anastomosis is not properly achieved, so that pancreatic juiceflows out through the gap, causing the pancreatic leakage.

Accordingly, recently, pancreas-jejunum anastomosis, pancreas-stomachanastomosis, pancreatic duct-mucosal anastomosis, dunking, stentimplantation and the like have been proposed to prevent the occurrenceof postoperative pancreatic leakage. In addition to these surgicalsuturing methods, somatostatin, fibrin glue or the like has also beenproposed.

The present inventors have found that novel compounds synthesized usingthe structure of existing antibiotics as a basic skeleton cantemporarily harden the pancreas, and thus can increase the efficiency ofsuturing during pancreatic surgery such as pancreaticoduodenectomy,thereby completing the present invention.

DISCLOSURE Technical Problem

The present inventors have developed novel compounds capable oftemporarily hardening the pancreas, making it possible to moreefficiently suture the pancreas in order to prevent pancreatic leakagefrom occurring after pancreaticoduodenectomy. As a result, the presentinventors have found that compound derivatives according to the presentinvention can temporarily increase the hardness and tension of softtissue such as the pancreas, thereby completing the present invention.

Therefore, it is an object of the present invention to provide a novelcompound represented by formula 1 or a pharmaceutically acceptable saltthereof.

Another object of the present invention is to provide a method forpreparing the compound.

Still another object of the present invention is to provide apharmaceutical composition for hardening soft tissue.

Still another object of the present invention is to provide apharmaceutical composition for hardening the pancreas.

Still another object of the present invention is to provide apharmaceutical composition for preventing pancreatic leakage.

Still another object of the present invention is to provide a method forscreening a compound for hardening soft tissue.

Technical Solution

Hereinafter, various embodiments described herein will be described withreference to figures. In the following description, numerous specificdetails are set forth, such as specific configurations, compositions,and processes, etc., in order to provide a thorough understanding of thepresent invention. However, certain embodiments may be practiced withoutone or more of these specific details, or in combination with otherknown methods and configurations. In other instances, known processesand preparation techniques have not been described in particular detailin order to not unnecessarily obscure the present invention. Referencethroughout this specification to “one embodiment” or “an embodiment”means that a particular feature, configuration, composition, orcharacteristic described in connection with the embodiment is includedin at least one embodiment of the present invention. Thus, theappearances of the phrase “in one embodiment” or “an embodiment” invarious places throughout this specification are not necessarilyreferring to the same embodiment of the present invention. Additionally,the particular features, configurations, compositions, orcharacteristics may be combined in any suitable manner in one or moreembodiments.

In accordance with one embodiment of the present invention, there isprovided a compound represented by the following formula 1 or apharmaceutically acceptable salt or hydrate thereof:

wherein

-   -   R is hydrogen or N(R′)(R″);    -   R′ and R″ are each independently hydrogen, a C₁₋₆ alkyl group or        a C₁₋₆ alkenyl group;    -   R₁ is hydrogen or a C₁₋₆ alkyl group;    -   the C₁₋₆ alkyl group of R₁ is unsubstituted or substituted with        R₂;    -   R₂ is selected from the group consisting of a C₁₋₆ alkenyl        group, a C₃₋₇ cycloalkyl group, a cyano group, a C₁₋₆ alkyl        group, a C₆₋₁₀ aryl group, —OR₃ and —COOR₄;    -   the aryl group of R₂ is unsubstituted or substituted with        halogen;    -   R₃ is —(CH₂)_(m)OR₅;    -   m is an integer ranging from 0 to 3; and    -   R₄ and R₅ are each independently a C₁₋₆ alkyl group.

In one embodiment of the present invention, the compound of formula 1 orthe pharmaceutically acceptable salt thereof may preferably be one ormore selected from among the compounds shown in Table 1 below.

TABLE 1 Structural formula Compound name Name

(2S,5R,6R)-3,3-dimethyl-7-oxo-6- (2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2- carboxylic acid —

Allyl (2S,5R,6R)-3,3-dimethyl-7- oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2- carboxylate KRM- 60028

Cyclohexylmethyl (2S,5R,6R)-3,3- dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate KRM-60029

3-cyanopropyl (2S,5R,6R)-3,3- dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate KRM-60030

3-(tert-butoxy)-3-oxopropyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylateKRM- 60077

Isobutyl (2S,5R,6R)-3,3-dimethyl- 7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2- carboxylate KRM- 60078

2-fluorobenzyl (2S,5R,6R)-3,3- dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate KRM-60079

2-(2-ethoxyethoxy)ethyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylateKRM- 60080

(2S,5R,6R)-6-((R)-2-amino-2- phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylic acid —

(2S,5R,6R)-cyclohexylmethyl 6- ((R)-2-amino-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate —

(2S,5R,6R)-3-cyanopropyl 6-((R)- 2-amino-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate —

(2S,5R,6R)-allyl 6-((R)-2-amino-2- phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate —

(2S,5R,6R)-allyl 6-((R)-2- (allylamino)-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate) —

In a preferred embodiment of the present invention, R′ and R″ may beeach independently hydrogen or a C₁₋₆ alkenyl group.

In a preferred embodiment of the present invention, R₁ may be hydrogenor a C₁₋₄ alkyl group.

In a preferred embodiment of the present invention, R₂ may be selectedfrom the group consisting of a C₁₋₃ alkenyl group, a C₅₋₇ cycloalkylgroup, a cyano group, a C₁₋₃ alkyl group, a C₆₋₈ aryl group, —OR₃ and—COOR₄.

In the present invention, the compound represented by formula 1 maypreferably be one or more selected from the group consisting of(2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylicacid; allyl(2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate;cyclohexylmethyl(2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate;3-cyanopropyl(2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylkate;and(2S,5R,6R)-6-((S)-2-amino-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylicacid.

The present invention also provides a pharmaceutically acceptable saltof the compound represented by formula 1. The pharmaceuticallyacceptable salt should have low toxicity to the human body and shouldnot affect the biological activity and physicochemical properties of itsparent compound. The pharmaceutically acceptable salt may be an acidaddition salt of the compound with a pharmaceutically acceptable freebase, but is not limited thereto.

The preferred salt forms of the compound according to the presentinvention may be, for example, salts with inorganic acids such ashydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate,phosphate, nitrate and carbonate; and salts with organic acids such asformic acid, acetic acid, propionic acid, oxalic acid, succinic acid,benzoic acid, citric acid, maleic acid, malonic acid, tartaric acid,gluconic acid, lactic acid, gentisic acid, fumaric acid, lactobionicacid, salicylic acid, trifluoroacetic acid and acetylsalicylic acid(aspirin); or salts with amino acids such as glycine, alanine, valine,isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine,arginine, tyrosine and proline; salts with sulfonic acids such asmethanesulfonate, ethanesulfonate, benzenesulfonate andtoluenesulfonate; metal salts formed by reaction with alkali metals suchas sodium and potassium; or salts with ammonium ions, but are notlimited thereto.

The salt can be prepared by a conventional method. For example, the saltcan be prepared by dissolving the compound of formula 1 in awater-miscible solvent, such as methanol, ethanol, acetone or1,4-dioxane, and then adding a free acid or a free base thereto,followed by crystallization.

In the present invention, a preferred example of the pharmaceuticallyacceptable salt of the compound represented by formula 1 may be one ormore selected from among the salts shown in Table 2 below.

TABLE 2 Structural formula Compound name Name

Sodium (2S,5R,6R)-6-((R)-2- amino-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylateKRM-60036

Potassium (2S,5R,6R)-3,3- dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylateKRM-60027

In addition, the hydrate form of the compound represented by formula 1may also be included within the scope of the present invention.

In accordance with another embodiment of the present invention, there isprovided a pharmaceutical composition for hardening soft tissue,comprising, as an active ingredient, the compound represented by formula1 or a pharmaceutically acceptable salt or hydrate thereof.

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

The compound represented by formula 1 and a pharmaceutically acceptablesalt and hydrate thereof, which are provided according to the presentinvention, can temporarily increase the hardness or tension of softtissue, thereby increasing the efficiency of suturing during suturing ofthe soft tissue, thereby preventing sequela or the like from occurringdue to insufficient anastomosis.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for preventing pancreaticleakage, comprising, as an active ingredient, the compound representedby formula 1 or a pharmaceutically acceptable salt or hydrate thereof.

The compound represented by formula 1 and a pharmaceutically acceptablesalt and hydrate thereof, which are provided according to the presentinvention, can temporarily increase the hardness or tension of thepancreas, thereby increasing the efficiency of suturing duringpancreaticoduodenectomy and effectively preventing pancreatic leakage.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for hardening soft tissue,comprising a step of administering to a subject an effective amount ofthe compound represented by formula 1 or a pharmaceutically acceptablesalt or hydrate thereof.

In the present invention, the subject may be a subject requiringtemporal hardening of soft tissue. More preferably, it may be a subjectrequiring surgical suturing of soft tissue.

In addition, in the present invention, the compound represented byformula 1 or a pharmaceutically acceptable salt or hydrate thereof maybe administered to the soft tissue of the subject.

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for preventing pancreaticleakage, comprising a step of administering to a subject an effectiveamount of the compound represented by formula 1 or a pharmaceuticallyacceptable salt or hydrate thereof.

In the present invention, the subject may be a subject requiringpancreaticoduodenectomy.

In addition, in the present invention, the compound represented byformula 1 or a pharmaceutically acceptable salt or hydrate thereof maybe administered to the pancreas of the subject.

The “effective amount” as used herein refers to an amount sufficient forachieving a temporal change in the hardness or tension of the softtissue or pancreas of the subject.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for hardening softtissue, comprising, as an active ingredient, a compound selected fromthe group consisting of a hydroxyquinoline compound, a derivativethereof, and a pharmaceutically acceptable salt and hydrate thereof.

In the present invention, the hydroxyquinoline compound and a derivativethereof may be one or more selected from among the compounds shown inTable 3 below.

TABLE 3 Structural formula Compound name Name

7-chloro-4-hydroxy-3- quinolinecarboxylic acid KRP-0001

5-chloro-8-hydroxyquinoline KRP-0002

5-chloro-8-hydroxy-7- iodoquinoline KRP-0003

5,7-diiodo-8-hydroxyquinoline KRP-0004

4,8-dihydroxyquinoline-2- carboxylic acid KRP-0005

4-hydroxy-2-methylquinoline KRP-0006

2-hydroxy-4-methylquinoline KRP-0007

8-hydroxy-5-nitroquinoline KRP-0008

4-hydroxy-7-trifluoromethyl-3- quinolinecarboxylic acid KRP-0009

3-hydroxyisoquinoline KRP-00010

4-hydroxy-7-methoxyquinoline KRP-00011

4-hydroxyquinoline KRP-00012

6-hydroxyquinoline KRP-00013

2-hydroxyquinoline KRP-00014

7-hydroxyquinoline KRP-00015

8-hydroxyquinoline-7- carboxylic acid KRP-00016

8-hydroxy-5-quinoline sulfonic acid —

8-hydroxyquinoline KRM-0042

In the present invention, the derivative of the hydroxyquinolinecompound may preferably be one or more selected from the groupconsisting of 7-chloro-4-hydroxy-3-quinolinecarboxylic acid;5-chloro-8-hydroxyquinoline; 5-chloro-8-hydroxy-7-iodoquinoline;5,7-diiodo-8-hydroxyquinoline; 4,8-dihydroxyquinoline-2-carboxylic acid;4-hydroxy-7-trifluoromethyl-3-quinolinecarboxylic acid;3-hydroxyisoquinoline; 4-hydroxyquinoline; 6-hydroxyquinoline;2-hydroxyquinoline; 8-hydroxyquinoline-7-carboxylic acid; and8-hydroxyquinoline. More preferably, it may be 3-hydroxyisoquinoline.

The present invention also provides a pharmaceutically acceptable saltof the hydroxyquinoline compound or the derivative of the compound. Thepharmaceutically acceptable salt should have low toxicity to the humanbody and should not affect the biological activity and physicochemicalproperties of its parent compound. The pharmaceutically acceptable saltmay be an acid addition salt of the basic compound with apharmaceutically acceptable free acid, but is not limited thereto.

The preferred salt forms of the compound according to the presentinvention may be, for example, salts with inorganic acids such ashydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate,phosphate, nitrate and carbonate; and salts with organic acids such asformic acid, acetic acid, propionic acid, oxalic acid, succinic acid,benzoic acid, citric acid, maleic acid, malonic acid, tartaric acid,gluconic acid, lactic acid, gentisic acid, fumaric acid, lactobionicacid, salicylic acid, trifluoroacetic acid and acetylsalicylic acid(aspirin); or salts with amino acids such as glycine, alanine, valine,isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine,arginine, tyrosine and proline; salts with sulfonic acids such asmethanesulfonate, ethanesulfonate, benzenesulfonate andtoluenesulfonate; metal salts formed by reaction with alkali metals suchas sodium and potassium; or salts with ammonium ions, but are notlimited thereto.

The salt can be prepared by a conventional method. For example, the saltcan be prepared by dissolving the hydroxyquinoline compound or thederivative of the compound in a water-miscible solvent, such asmethanol, ethanol, acetone or 1,4-dioxane, and then adding a free acidor a free base thereto, followed by crystallization.

In addition, the hydrate form of the hydroxyquinoline compound or thederivative of the compound may also be included within the scope of thepresent invention. Preferably, the hydrate form may be a hydrate shownin Table 4 below, but is not limited thereto.

TABLE 4 Structural formula Compound name Name

8-hydroxy-5-quinoline sulfonic acid hydrate KRP-0017

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

The hydroxyquinoline compound, derivative thereof and pharmaceuticallyacceptable salt and hydrate thereof according to the present inventioncan temporarily increase the hardness or tension of soft tissue, therebyincreasing the efficiency of suturing during suturing of the softtissue, thereby preventing sequela or the like from occurring due toinsufficient anastomosis.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for preventing pancreaticleakage, comprising, as an active ingredient, a compound selected fromthe group consisting of the hydroxyquinoline compound, a derivativethereof, and a pharmaceutically acceptable salt and hydrate thereof.

In the present invention, the hydroxyquinoline compound and thederivative thereof may be one or more selected from among the compoundsshown in Table 3 above.

In the present invention, the derivative of the hydroxyquinolinecompound may preferably be one or more selected from the groupconsisting of 5-hydroxyquinoline;7-chloro-4-hydroxy-3-quinolinecarboxylic acid;5-chloro-8-hydroxyquinoline; 5-chloro-8-hydroxy-7-iodoquinoline;5,7-diiodo-8-hydroxyquinoline; 4,8-dihydroxyquinoline-2-carboxylic acid;4-hydroxy-7-trifluoromethyl-3-quinolinecarboxylic acid;3-hydroxyisoquinoline; 4-hydroxyquinoline; 6-hydroxyquinoline;2-hydroxyquinoline; 8-hydroxyquinoline-7-carboxylic acid; and8-hydroxyquinoline. More preferably, it may be 3-hydroxyisoquinoline.

The present invention also provides a pharmaceutically acceptable saltof the hydroxyquinoline compound or the derivative of the compound. Thepharmaceutically acceptable salt should have low toxicity to the humanbody and should not affect the biological activity and physicochemicalproperties of its parent compound. The pharmaceutically acceptable saltmay be an acid addition salt of the basic compound with apharmaceutically acceptable free acid, but is not limited thereto.

The preferred salt forms of the compound according to the presentinvention may be, for example, salts with inorganic acids such ashydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate,phosphate, nitrate and carbonate; and salts with organic acids such asformic acid, acetic acid, propionic acid, oxalic acid, succinic acid,benzoic acid, citric acid, maleic acid, malonic acid, tartaric acid,gluconic acid, lactic acid, gentisic acid, fumaric acid, lactobionicacid, salicylic acid, trifluoroacetic acid and acetylsalicylic acid(aspirin); or salts with amino acids such as glycine, alanine, valine,isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine,arginine, tyrosine and proline; salts with sulfonic acids such asmethanesulfonate, ethanesulfonate, benzenesulfonate andtoluenesulfonate; metal salts formed by reaction with alkali metals suchas sodium and potassium; or salts with ammonium ions, but are notlimited thereto.

The salt can be prepared by a conventional method. For example, the saltcan be prepared by dissolving the hydroxyquinoline compound or thederivative of the compound in a water-miscible solvent, such asmethanol, ethanol, acetone or 1,4-dioxane, and then adding a free acidor a free base thereto, followed by crystallization.

In addition, the hydrate form of the hydroxyquinoline compound or thederivative of the compound may also be included within the scope of thepresent invention. Preferably, the hydrate form may be the hydrate shownin Table 4 above, but is not limited thereto.

The hydroxyquinoline compound, derivative thereof and pharmaceuticallyacceptable salt and hydrate thereof according to the present inventioncan temporarily increase the hardness or tension of the pancreas,thereby increasing the efficiency of suturing duringpancreaticoduodenectomy and effectively preventing pancreatic leakage.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for hardening soft tissue,comprising a step of administering to a subject an effective amount of acompound selected from the group consisting of the hydroxyquinolinecompound, a derivative thereof, and a pharmaceutically acceptable saltand hydrate thereof.

In the present invention, the hydroxyquinoline compound, a derivativethereof, and a pharmaceutically acceptable salt and hydrate thereof areas described above with respect to the pharmaceutical composition forhardening soft tissue according to the present invention, and thus thedetailed description thereof will be omitted herein.

In the present invention, the subject may be a subject requiringtemporal hardening of soft tissue. More preferably, it may be a subjectrequiring surgical suturing of soft tissue.

In addition, in the present invention, the hydroxyquinoline compound, aderivative thereof, or a pharmaceutically acceptable salt or hydratethereof may be administered to the soft tissue of the subject.

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for preventing pancreaticleakage, comprising a step of administering to a subject an effectiveamount of a compound selected from the group consisting ofhydroxyquinoline compound, a derivative thereof, and a pharmaceuticallyacceptable salt and hydrate thereof.

In the present invention, the hydroxyquinoline compound, a derivativethereof, and a pharmaceutically acceptable salt and hydrate thereof areas described above with respect to the pharmaceutical composition forpreventing pancreatic leakage according to the present invention, andthus the detailed description thereof will be omitted herein.

In the present invention, the subject may be a subject requiringpancreaticoduodenectomy.

In addition, in the present invention, the hydroxyquinoline compound, aderivative thereof, or a pharmaceutically acceptable salt or hydratethereof may be administered to the pancreas of the subject.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for hardening softtissue, comprising, as an active ingredient, a compound represented byany one of the following formulas 2 to 9, or a pharmaceuticallyacceptable salt or hydrate thereof:

In the present invention, the detailed names of the compoundsrepresented by formulas 2 to 9 are as shown in Table 5 below.

TABLE 5 Structural formula Compound name Name

(S)-2-amino-3-mercapto-3- methylbutanoic acid KRM-60037

1-(4-(4-(((2R,4S)-2-((1H- imidazol-1-yl)methyl)-2-(2,4-dichlorophenyl)-1,3-dioxolan-4- yl)methoxy)phenyl)piperazin-1-yl)ethanone KRM-60049

(6R,7R)-3-(acetoxymethyl)-7- amino-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid KRM-60081

8-ethyl-5-oxo-2-(piperazin-1-yl)- 5,8-dihydropyrido[2,3-d]pyrimidine-6-carboxylic acid KRM-60082

Sodium (2S,3S,5R)-3-((1H-1,2,3- triazol-1-yl)methyl)-3-methyl-7-oxo-4-thia-1- azabicyclo[3.2.0]heptane-2- carboxylate 4,4-dioxideKRM-60084

Sodium (6R,7R)-3- (acetoxymethyl)-7-((Z)-2-(2- aminothiazol-4-yl)-2-(methoxyimino)acetamido)-8- oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate KRM-60086

(6R,7R)-3-(acetoxymethyl)-8- oxo-7-(2-(thiophen-2-yl)acetamido)-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2- carboxylic acid —

5-(3,4,5- trimethoxybenzyl)pyrimidine-2,4- diamine KRM-60088

The present invention also provides pharmaceutically acceptable salts ofthe compounds represented by formulas 2 to 9. The pharmaceuticallyacceptable salts should have low toxicity to the human body and shouldnot affect the biological activities and physicochemical properties oftheir parent compounds. The pharmaceutically acceptable salts may beacid addition salts of the compounds of formulas 2 to 9 withpharmaceutically acceptable bases, but are not limited thereto.

The preferred salt forms of the compounds according to the presentinvention may be, for example, salts with inorganic acids such ashydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate,phosphate, nitrate and carbonate; and salts with organic acids such asformic acid, acetic acid, propionic acid, oxalic acid, succinic acid,benzoic acid, citric acid, maleic acid, malonic acid, tartaric acid,gluconic acid, lactic acid, gentisic acid, fumaric acid, lactobionicacid, salicylic acid, trifluoroacetic acid and acetylsalicylic acid(aspirin); or salts with amino acids such as glycine, alanine, valine,isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine,arginine, tyrosine and proline; salts with sulfonic acids such asmethanesulfonate, ethanesulfonate, benzenesulfonate andtoluenesulfonate; metal salts formed by reaction with alkali metals suchas sodium and potassium; or salts with ammonium ions, but are notlimited thereto.

The salt can be prepared by a conventional method. For example, the saltcan be prepared by dissolving the compound represented by any one offormulas 2 to 9 in a water-miscible solvent, such as methanol, ethanol,acetone or 1,4-dioxane, and then adding a free acid or a free basethereto, followed by crystallization.

In the present invention, a preferred example of a pharmaceuticallyacceptable salt of the compound represented by formula 8 may be the saltshown in Table 6 below.

TABLE 6 Structural formula Compound name Name

Sodium (6R,7R)-3- (acetoxymethyl)-8-oxo-7-(2-(thiophen-2-yl)acetamido)-5- thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate KRM-60087

In addition, the hydrate forms of the compounds represented by formulas2 to 9 may also be included within the scope of the present invention.

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

The compounds represented by formulas 2 to 9 and pharmaceuticallyacceptable salts and hydrates thereof according to the present inventioncan temporarily increase the hardness or tension of soft tissue, therebyincreasing the efficiency of suturing during suturing of the softtissue, thereby preventing sequela or the like from occurring due toinsufficient anastomosis.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for preventing pancreaticleakage, comprising, as an active ingredient, a compound represented byany one of the following formulas 2 to 9, or a pharmaceuticallyacceptable salt or hydrate thereof:

The present invention also provides pharmaceutically acceptable salts ofthe compounds represented by formulas 2 to 9. The pharmaceuticallyacceptable salts should have low toxicity to the human body and shouldnot affect the biological activities and physicochemical properties oftheir parent compounds. The pharmaceutically acceptable salts may beacid addition salts of the compounds of formulas 2 to 9 withpharmaceutically acceptable free bases, but are not limited thereto.

The preferred salt forms of the compounds according to the presentinvention may be, for example, salts with inorganic acids such ashydrochloric acid, bromic acid, sulfuric acid, sodium hydrogen sulfate,phosphate, nitrate and carbonate; and salts with organic acids such asformic acid, acetic acid, propionic acid, oxalic acid, succinic acid,benzoic acid, citric acid, maleic acid, malonic acid, tartaric acid,gluconic acid, lactic acid, gentisic acid, fumaric acid, lactobionicacid, salicylic acid, trifluoroacetic acid and acetylsalicylic acid(aspirin); or salts with amino acids such as glycine, alanine, valine,isoleucine, serine, cysteine, cystine, aspartic acid, glutamine, lysine,arginine, tyrosine and proline; salts with sulfonic acids such asmethanesulfonate, ethanesulfonate, benzenesulfonate andtoluenesulfonate; metal salts formed by reaction with alkali metals suchas sodium and potassium; or salts with ammonium ions, but are notlimited thereto.

The salt can be prepared by a conventional method. For example, the saltcan be prepared by dissolving the compound represented by any one offormulas 2 to 9 in a water-miscible solvent, such as methanol, ethanol,acetone or 1,4-dioxane, and then adding a free acid or a free basethereto, followed by crystallization.

In the present invention, a preferred example of a pharmaceuticallyacceptable salt of the compound represented by formula 8 may be the saltshown in Table 6 above.

In addition, the hydrate forms of the compounds represented by formulas2 to 9 may also be included within the scope of the present invention.

The compounds represented by formulas 2 to 9 or pharmaceuticallyacceptable salts and hydrates thereof according to the present inventioncan temporarily increase the hardness or tension of pancreas, therebyincreasing the efficiency of suturing during pancreaticoduodenectomy andeffectively preventing pancreatic leakage.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for hardening soft tissue,comprising a step of administering to a subject an effective amount ofthe compound represented by any one of formulas 2 to 9 or apharmaceutically acceptable salt or hydrate thereof.

In the present invention, the compounds represented by formulas 2 to 9and pharmaceutically acceptable salts and hydrates thereof are asdescribed above with respect to the pharmaceutical composition forhardening soft tissue according to the present invention, and thus thedetailed description thereof will be omitted herein.

In the present invention, the subject may be a subject requiringtemporal hardening of soft tissue. More preferably, it may be a subjectrequiring surgical suturing of soft tissue.

In addition, in the present invention the compound represented by anyone of formulas 2 to 9 or a pharmaceutically acceptable salt or hydratethereof may be administered to the soft tissue of the subject.

In the present invention, the “soft tissue” may be one or more tissuesselected from the group consisting of pancreas, liver, nerve, ligament,serosa, myofascia, intervertebral disc, and blood vessel, and maypreferably be pancreas, but is not limited thereto.

In accordance with still another embodiment of the present invention,the present invention is directed to a method for preventing pancreaticleakage, comprising a step of administering to a subject an effectiveamount of the compound represented by any one of formulas 2 to 9 or apharmaceutically acceptable salt or hydrate thereof.

In the present invention, the compounds represented by formulas 2 to 9or pharmaceutically acceptable salts or hydrates thereof are asdescribed above with respect to the pharmaceutical composition forpreventing pancreatic leakage according to the present invention, andthus the detailed description thereof will be omitted herein.

In the present invention, the subject may be a subject requiringpancreaticoduodenectomy.

In addition, in the present invention, the compounds represented byformulas 2 to 9 or pharmaceutically acceptable salts or hydrates thereofmay be administered to the pancreas of the subject.

In accordance with still another embodiment of the present invention,there is provided a method for preparing a compound derivative,comprising the steps of: dissolving a compound represented by thefollowing formula 10 and18-crown-6(1,4,7,10,13,16-hexaoxacyclooctadecane) in an organic solvent;and

adding a compound represented by the following formula 11 to thedissolved solution, followed by stirring, thereby synthesizing acompound:

R₆—Br  [Formula 11]

In formula 10 above,

R is hydrogen or N(R′)(R″), and

R′ and R″ are each independently hydrogen, a C₁₋₆ alkyl group or a C₁₋₆alkenyl group, and

in formula 11 above,

R₆ is hydrogen or a C₁₋₆ alkyl group;

the C₁₋₆ alkyl group of R₆ is unsubstituted or substituted with R₇;

R₇ is selected from the group consisting of a C₁₋₆ alkenyl group, a C₃₋₇cycloalkyl group, a cyano group, a C₁₋₆ alkyl group, a C₆₋₁₀ aryl group,—OR₈ and —COOR₉;

the aryl group of R₇ is unsubstituted or substituted with halogen;

R₈ is —(CH₂)_(n)OR₁₀;

n is an integer ranging from 0 to 3;

R₉ and R₁₀ are each independently C₁₋₆ alkyl.

In the present invention, the compound represented by formula 11 may beselected from the group consisting of 3-bromopropene,(bromomethyl)cyclohexane, 4-bromobutyronitrile, tert-butylbromopropionate, 1-bromo-2-methylpropane, 2-fluorobenzyl bromide, and2-(2-ethoxyethoxy)ethyl bromide, but is not limited as long as it is analkyl bromide that may be substituted by reaction with the carboxylateof the formula.

In the present invention, the organic solvent is used to dissolve thecompounds, and is preferably an aprotic polar solvent. Specifically, asthe solvent, dioxane, tetrahydrofuran (THF), acetone, dimethyl suifoxide(DMSO), dimethylformamide (DIVLF), 1-methyl-2-pyrrolidone (NMP) or thelike may be used. More preferably, dimethylformamide (DMF) is used.

In accordance with still another embodiment of the present invention,there is provided a method for screening a compound for hardening thepancreas, the method comprising the steps of: (a) measuring the hardnessor tension of the pancreas of a non-human mammal not treated with acandidate compound; (b) measuring the hardness or tension of thepancreas of a non-human mammal treated with the candidate compound; and(c) determining that when the hardness or tension of the pancreas,measured in step (b), is higher than the hardness or tension of thepancreas, measured in step (a), the compound is the compound forhardening the pancreas.

In the present invention, the KRM-60027 (penicillin G potassium salt)and KRM-60036 (ampicillin sodium salt) compounds are antibiotics having(R)-4-thia-1-azabicyclo[3,2,0]hepta-7-one as a skeleton, and belong tothe same family. The two compounds differ in that KRM-60027 is apotassium salt and KRM-60036 is a sodium salt. In addition, KRM-60027has a CH₂-bond at the benzylic position on the left side of theskeleton, whereas KRM-60036 has an amino acid substitution for one ofhydrogen atoms present in the CH₂-bond at the benzylic position, whichhas the effect of increasing the pharmacokinetics (PK) of the drug.KRM-60027 showed the result of significantly increasing the tension ofthe pancreas, and KRM-60036 was found to significantly increase thehardness of the pancreas.

In the present invention, KRM-60037 (D-penicillamine) is a drug having asimple structure. It has a molecular weight of only 149, but exhibitedthe effect of increasing the tension by 1.5 times. In particular, thissmall molecule has advantages in that it has chemically modifiablemoieties, and thus various substituents can be attached to the molecule,thereby sufficiently increasing the tension and hardness of thepancreas.

In the present invention, KRM-60042 (8-hydroxyquinoline) is also alow-molecular-weight compound having a simple skeleton and a molecularweight of 145, and various substituents can be easily attached thereto.It can be seen that KRM-60042 significantly increases both the hardnessand tension of the pancreas.

In the present invention, KRM-60049 (ketoconazole) has advantages inthat it has an ideal molecular weight of 530 and contains manyfunctional groups capable of forming salts such as piperazine andimidazole salts, thus making various salts.

In the present invention, the “hardness” means the level of resistanceto the strain of particularly soft tissue, preferably the pancreas,among human organs, when an external force is applied to the outside ofthe pancreas. The measurement of the hardness according to the presentinvention is performed using a hardness meter, and measurement of thehardness of the pancreas is performed by measuring elastic modulus (EM)based on the Hooke's law and the Hertz's contact stress theory. The EMis a measure of the stiffness of an elastic material, and is expressedas the ratio of “the stress along the axis” and “the total strain alongthe axis within the range of the stress” by the Hooke's law.

In the present invention, the “tension” means a force which occurs inparticularly soft tissue, preferably the pancreas, among human organs.Specifically, it means a force which occurs in an organ in response toan external force exerted by stretching or pulling action. Themeasurement of the tension according to the present invention isperformed using a dynamometer. More specifically, the tension isdetermined by measuring suture-holding capacity using a Newtondynamometer. At this time, the tension is calculated by adding theweight of the tissue.

In one embodiment of the present invention, the “alkyl” means saturatedstraight or branched chain hydrocarbon groups, for example, methyl,ethyl, propyl, butyl, isobutyl, tert-butyl, and pentyl. As used herein,the “C₁₋₆ alkyl group” means an alkyl group containing 1 to 6 carbonatoms.

In one embodiment of the present invention, the “cycloalkyl” means amonocyclic or polycyclic saturated ring containing carbon and hydrogenatoms.

In one embodiment of the present invention, the “aryl” means a fully orpartially unsaturated monocyclic or polycyclic carbon ring havingaromaticity. The aryl group of the present invention is preferablymonocryl or biaryl.

In one embodiment of the present invention, the “alkenyl” means an alkylradical having one or more carbon-carbon double bonds. Here, the alkylis as defined above.

In one embodiment of the present invention, the “halogen” or “halo”means fluorine, chlorine, bromine or iodine, unless otherwise indicated.

The pharmaceutical composition of the present invention comprises apharmaceutically acceptable carrier in addition to the active ingredientcompound. The pharmaceutically acceptable carrier that is contained inthe pharmaceutical composition of the present invention is one that isgenerally used in drug formulations, and examples thereof include, butare not limited to, lactose, dextrose, sucrose, sorbitol, mannitol,starch, acacia rubber, potassium phosphate, alginate, gelatin, potassiumsilicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose,water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

The pharmaceutical composition according to the present invention mayfurther comprise lubricants, wetting agents, sweeteners, flavoringagents, emulsifiers, suspending agents, and preservatives. Details ofsuitable pharmaceutically acceptable carriers and formulations can befound in Remington's Pharmaceutical Sciences (19^(th) edition, 1995).

The pharmaceutical composition according to the present invention may beadministered orally or parenterally, preferably parenterally. Parenteraladministration may be performed by intravenous injection, subcutaneousinjection, intramuscular injection, intraperitoneal injection,transdermal administration or intra-articular injection. Morespecifically, it is performed by intramuscular injection orintraperitoneal injection.

The suitable dose of the pharmaceutical composition of the presentinvention may vary depending on the drug formulation method, the mode ofadministration, the patient's age, body weight, sex, disease condition,diet, the period of administration, the route of administration, therate of excretion, and sensitivity to the pharmaceutical compositionused. Preferably, the pharmaceutical composition of the presentinvention may be administered at a daily dose of 0.001 to 10000 mg/kg(body weight).

According to conventional techniques known to those skilled in the art,the pharmaceutical composition according to the present invention isformulated with a pharmaceutically acceptable carrier and/or vehicle asdescribed above, finally providing several forms including a unit doseform and a multi-dose form. Non-limiting examples of the formulationsinclude, but not limited to, a solution, a suspension or an emulsion inoil or aqueous medium, an elixir, a powder, a granule, a tablet and acapsule, and may further comprise a dispersion agent or a stabilizer.

Advantageous Effects

The compounds according to the present invention can temporarilyincrease the hardness or tension of soft tissue, thereby increasing theefficiency of suturing during suturing of the soft tissue, therebypreventing sequela or the like from occurring due to insufficientanastomosis.

In addition, the compounds according to the present invention cantemporarily increase the hardness or tension of soft tissue,particularly the pancreas, thereby increasing the efficiency of suturingduring pancreaticoduodenectomy and effectively preventing pancreaticleakage.

DESCRIPTION OF DRAWINGS

FIG. 1 graphically shows the changes in the hardness of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60027,KRM-60036, KRM-60037, KRM-60042 and KRM-60049 according to the presentinvention in one example of the present invention.

FIG. 2 graphically shows the changes in the tension of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60027,KRM-60036, KRM-60037, KRM-60042 and KRM-60049 according to the presentinvention in one example of the present invention;

FIG. 3 graphically shows the changes in the hardness of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60028,KRM-60029 and KRM-60030 according to the present invention in oneexample of the present invention.

FIG. 4 graphically shows the changes in the tension of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60028,KRM-60029 and KRM-60030 according to the present invention in oneexample of the present invention.

FIG. 5 graphically shows the changes in the hardness of the mousepancreas after treatment of the pancreas with the antibiotics ICRP-00001to KRP-000017 according to the present invention in one example of thepresent invention.

FIG. 6 graphically shows the changes in the tension of the mousepancreas after treatment of the pancreas with the antibiotics KRP-00001to KRP-00017 according to the present invention in one example of thepresent invention.

FIG. 7 graphically shows the changes in the hardness of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60077to KRM-60088 according to the present invention in one example of thepresent invention.

FIG. 8 graphically shows the changes in the tension of the mousepancreas after treatment of the pancreas with the antibiotics KRM-60077to KRM-60088 according to the present invention in one example of thepresent invention.

BEST MODE

In accordance with one embodiment of the present invention, there isprovided a compound represented by the following formula 1 or apharmaceutically acceptable salt or hydrate thereof:

wherein

-   -   R is hydrogen or N(R′)(R″);    -   R′ and R″ are each independently hydrogen, a C₁₋₆ alkyl group or        a C₁₋₆ alkenyl group;    -   R₁ is hydrogen or a C₁₋₆ alkyl group;    -   the C₁₋₆ alkyl group of R₁ is unsubstituted or substituted with        R₂;    -   R₂ is selected from the group consisting of a C₁₋₆ alkenyl        group, a C₃₋₇ cycloalkyl group, a cyano group, a C₁₋₆ alkyl        group, a C₆₋₁₀ aryl group, —OR₃ and —COOR₄;    -   the aryl group of R₂ is unsubstituted or substituted with        halogen;    -   R₃ is —(CH₂)_(m)OR₅;    -   m is an integer ranging from 0 to 3; and    -   R₄ and R₅ are each independently a C₁₋₆ alkyl group.

In accordance with another embodiment of the present invention, there isprovided a pharmaceutical composition for hardening soft tissue,comprising, as an active ingredient, the compound represented by formula1 or a pharmaceutically acceptable salt or hydrate thereof.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for hardening softtissue, comprising, as an active ingredient, a compound selected fromthe group consisting of a hydroxyquinoline compound, a derivativethereof, and a pharmaceutically acceptable salt and hydrate thereof.

In accordance with still another embodiment of the present invention,there is provided a pharmaceutical composition for hardening softtissue, comprising, as an active ingredient, a compound represented byany one of the following formulas 2 to 9 or a pharmaceuticallyacceptable salt or hydrate thereof:

MODE FOR INVENTION

Hereinafter, the present invention will be described in detail withreference to examples. It will be obvious to those skilled in the artthat these examples are only intended to illustrate the presentinvention in more detail and that the scope of the present invention asdefined by the appended claims is not limited by these examples.

Example 1 Selection of Starting Compounds for Synthesis of Compounds forHardening Pancreas

To select starting compounds for synthesizing compounds that can enhancethe function of the pancreas, a total of 23 antibiotics as shown belowwere prepared:

More specifically, three mice were treated with 1 mM of each of theantibiotics shown above, and three mice were treated with phosphatebuffered saline (PBS) as a control. Furthermore, the tension andhardness of the pancreases of the mice treated with each antibiotic werecompared with those of the pancreases of the control mice not treatedwith the antibiotic, and antibiotics showing increased tension andhardness upon treatment were selected as starting compounds.

As a result, it was confirmed that among the 23 compounds, onlyKRM-60027, KRM-60036, KRM-60037, KRM-60042 and KRM-60049 compounds hadthe effects of increasing the tension and hardness of the pancreas.FIGS. 1 and 2 graphically show the changes in the hardness and tensionof the pancreas after treatment with these five compounds compared tothe control. Here, the hardness and the tension were measured using themethods described below.

(1) Measurement of Hardness

In order to measure the resistance of the pancreas using a hardnessmeter, the elastic modulus (EM) of the dissected pancreas was measuredusing Venustron (Axiom, Koriyama, Japan) based on the Hooke's law andthe Hertz's contract stress theory, and the resistance of the pancreaswas calculated using the following equation:

$E = {\frac{3}{4} \cdot F \cdot \left( {1 - \upsilon^{2}} \right) \cdot t^{- \frac{3}{2}} \cdot {R^{- \frac{1}{2}}.}}$

(F, force; v, Poisson's ratio; t, variant of compression; r, thediameter of a rigid spherical probe that presses an object vertically)

(2) Measurement of Tensile

The tension of the pancreas was measured using a dynamometer.Specifically, a pancreatic fragment having a size of 1×1×1.5 cm wasprepared, and then silk 4-0 suture was passed through the tissue to forma knob, and suture-holding capacity was measured using a Newtondynamometer. At this time, the tension was calculated by adding theweight of the tissue.

As shown in FIGS. 1 and 2 , it could be seen that the pancreas of themice was treated with each of the five antibiotics according to thepresent invention, that is, KRM-60027 (penicillin G potassium salt),KRM-60036 (ampicillin sodium salt), KRM-60037 (D-penicillamine),KRM-60042 (8-hydroxyquinoline) and KRM-60049 (ketoconazole), the tensionand/or hardness of the pancreas significantly increased compared to thecontrol. More specifically, it could be seen that KRM-60027 increasedthe tension of the pancreas to a significant level, and KRM-60036significantly increased the hardness of the pancreas. In addition, itcould be seen that KRM-60037, a drug having a simple structure and amolecular weight of only 149, exhibited the effect of increasing thetension of the pancreas by 1.5 times, and KRM-60042 and KRM-60049increased both the hardness and tension of the pancreas to high levels.

Based on these results, the five antibiotics were selected as startingcompounds for compounds capable of hardening the pancreas.

Synthetic Example 1 Synthesis of Derivatives of KRM-60027

The KRM-60027 compound which showed an excellent effect on an increasein the tension of the pancreas in Example 1 above was used as a startingmaterial. Various alkyl bromide compounds were bound at the carboxylateposition of KRM-60027, thereby synthesizing final compounds forenhancing the function of the pancreas.

Synthetic Example 1-1 Synthesis of KRM-60028

The KRM-60027 compound used as a starting material and 18-crown-6(1,4,7,10,13,16-hexaoxacyclooctadecane) were dissolved in DMF(dimethylformamide), and then 3-bromopropene as alkyl bromide was addedthereto, followed by stirring at room temperature for 24 hours. Aftercompletion of the reaction was confirmed by thin-layer chromatography(TLC), ethyl acetate was added to the reaction solution which was thenwashed several times with water. The ethyl acetate layer containing theproduct was separated, and then anhydrous sodium sulfate was addedthereto to remove the remaining water, followed by concentration. Theconcentrate was purified by column chromatography, thereby obtaining thefinal product (KRM-60028) in which the carboxylate of KRM-60027 wassubstituted with allyl.

Synthetic Example 1-2 Synthesis of KRM-60029

A final product in which the carboxylate of KRM-60027 was substitutedwith cyclohexyl was obtained in the same manner as described inSynthetic Example 1-1, except that 4-(bromomethyl)cyclohexane was usedinstead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-3 Synthesis of KRM-60030

A final product in which the carboxylate of KRM-60027 was substitutedwith cyanobutyl was obtained in the same manner as described inSynthetic Example 1-1, except that 4-bromobutyronitrile was used insteadof 3-bromopropene as the alkyl bromide.

Synthetic Example 1-4 Synthesis of KRM-60077

A final product in which the carboxylate of KRM-60027 was substitutedwith tert-butyl propionate was obtained in the same manner as describedin Synthetic Example 1-1, except that tert-butyl bromopropionate wasused instead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-5 Synthesis of KRM-60078

A final product in which the carboxylate of KRM-60027 was substitutedwith isobutyl was obtained in the same manner as described in SyntheticExample 1-1, except that 1-bromo-2-methylpropane was used instead of3-bromopropene as the alkyl bromide.

Synthetic Example 1-6 Synthesis of KRM-60079

A final product in which the carboxylate of KRM-60027 was substitutedwith 2-fluorobenzyl was obtained in the same manner as described inSynthetic Example 1-1, except that 2-fluorobenzyl bromide was usedinstead of 3-bromopropene as the alkyl bromide.

Synthetic Example 1-7 Synthesis of KRM-60080

A final product in which the carboxylate of KRM-60027 was substitutedwith ethylene glycol was obtained in the same manner as described inSynthetic Example 1-1, except that 2-(2-ethoxyethoxy)ethyl bromide wasused instead of 3-bromopropene as the alkyl bromide.

Example 2 Additional Selection of Compounds for Hardening Pancreas

The pancreas of mice was treated with each of KRM-60028, KRM-60029 andKRM-60030 (synthesized in Synthetic Example 1 above) in the same manneras described in Example 1 above, and then changes in the hardness andtension of the pancreas compared to those of an untreated control groupwere measured. The results of the measurement are graphically shown inFIGS. 3 and 4 .

As shown in FIGS. 3 and 4 , it could be confirmed that when the pancreasof mice was treated with each of KRM-60028, KRM-60029 and KRM-60030according to the present invention, both the hardness and tension of thepancreas increased compared to those of the control. Referring toExample 1, the starting compound KRM-60027 showed an insignificantchange in the hardness of the pancreas and a significant increase in thetension of the pancreas, compared to the control. It could be seen thatKRM-60028, KRM-60029 and KRM-60030, which are compounds modified fromKRM-60027, increased both the hardness and tension of the pancreas. Inparticular, the KRM-60030 compound is a derivative obtained by attachinga cyanopropyl group to the carboxylate group of KRM-60027, and has anitrile group at the right end portion, and thus has a functional groupcapable of functioning as a hydrogen bonding acceptor. It could be seenthat this KRM-60030 compound increase the hardness of the pancreas by1.5 times and the tension of the pancreas by 2 times, compared to thoseof the control.

This suggests that the compound represented by formula 1 according tothe present invention can increase the hardness and tension of thepancreas, thereby temporarily hardening the pancreas.

Synthetic Example 2 Synthesis of Derivatives of KRM-60036

The KRM-60036 compound which showed an excellent effect on an increasein the hardness of the pancreas in Example 1 above was used as astarting material. Alkyl bromide compounds were introduced selectivelyat the carboxylate position of KRM-60036 while maintaining the amine ofKRM-60036, thereby synthesizing final compounds for enhancing thefunction of the pancreas. Specifically, as shown in reaction scheme 1below, the amine of the KRM-60036 compound used as a starting materialwas subjected to boc protection. Then, as shown in reaction scheme 2below, using 3-bromopropene, (bromomethyl)cyclohexane and4-bromobutyronitrile as alkyl bromides, an alkyl group was introduced inthe same manner as described in Synthetic Example 1-1 above, followed byboc deprotection, thereby obtaining final amine compounds.

Example 3 Derivatives of KRM-60042

The following derivative compounds were prepared, which are based on ahydroxyl group and quinoline which are the basic skeleton of theKRM-60042 compound (Sigma-Aldrich, cat. No 252565 148-24-3) which showedexcellent effects on increases in both the hardness and tension of thepancreas in Example 1 above.

The pancreas of mice was treated with each of KRP-0001 to KRP-00017 (atotal of 17 compounds) in the same manner as described in Example 1above, and then the changes in the hardness and tension of the pancreascompared to those of an untreated control group were measured. Theresults of the measurement are graphically shown in FIGS. 5 and 6 .

As shown in FIGS. 5 and 6 , it could be confirmed that among the 17derivatives having the quinoline of KRM-60042 as a skeleton, treatmentwith 11 compounds (KRP-00001, KRP-00002, KRP-00003, KRP-00004,KRP-00005, KRP-00006, KRP-00009, KRP-00012, KRP-00013, KRP-00014 andKRP-00016) increased both the hardness and tension of the pancreas, andparticularly, treatment with 10 compounds (ICRP-00001, KRP-00002,KRP-00003, KRP-00004, KRP-00005, KRP-00009, KRP-00012, KRP-00013,KRP-00014 and KRP-00016) increased the tension by at least 1.5 times,and treatment with KRP-00010 increased the tension by about 2.4 times,compared to that of the control group.

This suggests that the hydroxyquinoline derivatives according to thepresent invention exhibited the effect of significantly increasing thetension of the pancreas and also increased the hardness of the pancreas.

Example 4 Other Compounds

The pancreas of mice was treated with each of KRM-60077 to KRM-60080(synthesized in Synthetic Example 1) and KRM-60081 to KRM-60088compounds (shown below) in the same manner as described in Example 1above, and then the changes in the hardness and tension of the pancreascompared to those of an untreated control group were measured. Theresults of the measurement are graphically shown in FIGS. 7 and 8 .

As shown in FIGS. 7 and 8 , it could be confirmed that treatment withthe KRM-60077 to KRM-60088 compounds increased the hardness and/or thetension of the pancreas compared to those of the control group, andparticularly, treatment with the KRM-60077, KRM-60078, KRM-60080,KRM-60081, KRM-60082 and KRM-60088 compounds increased both the hardnessand tension of the pancreas to significant levels.

This suggests that the compounds according to the present inventionexhibited the effect of significantly increasing the tension of thepancreas and also increased the hardness of the pancreas.

The above experimental results suggest that when various modifiedcompounds (derivatives) based on the structures of antibiotics thatincrease the tension and/or hardness of the pancreas are synthesizedaccording to the present invention, compounds that effectively increasethe function of the pancreas can be obtained. In addition, it can beseen that when the pancreas is treated with the compounds of the presentinvention, obtained as described above, the hardness and tension of thepancreas can temporarily increase, and thus the efficiency of suturingof the pancreas in surgical operations such as pancreaticoduodenectomycan increase, and furthermore, pancreatic leakage can also be prevented.

INDUSTRIAL APPLICABILITY

The present invention relates to compounds capable to temporarilyhardening soft tissue for surgical suturing of the soft tissue.

The invention claimed is:
 1. A compound or a pharmaceutically acceptable salt or hydrate thereof, wherein the compound is selected from the group consisting of the following compounds: 1) Cyclohexylmethyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; 2) 3-(tert-butoxy)-3-oxopropyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; 3) 2-fluorobenzyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate; and 4) 2-(2-ethoxyethoxy)ethyl (2S,5R,6R)-3,3-dimethyl-7-oxo-6-(2-phenylacetamido)-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate.
 2. A pharmaceutical composition for hardening soft tissue, comprising, as an active ingredient a compound according to claim
 1. 3. The pharmaceutical composition of claim 2, wherein the soft tissue is one or more tissues selected from the group consisting of pancreas, liver, nerve, ligament, serosa, myofascia, intervertebral disc, and blood vessel.
 4. A pharmaceutical composition for preventing pancreatic leakage, comprising, as an active ingredient a compound according to claim
 1. 5. A method for preparing a compound according to claim 1, comprising the steps of: dissolving a compound represented by the following formula 10 and 18-crown-6 (1,4,7,10,13,16-hexaoxacyclooctadecane) in an organic solvent; and adding an alkyl bromide to the dissolved solution, followed by stirring, thereby synthesizing a compound of claim 1:

wherein R is hydrogen or N(R′)(R″), and R′ and R″ are each independently hydrogen or a Ci-6 alkenyl group wherein an alkyl bromide is one or more selected from the group consisting of (bromomethyl)cyclohexane, tert-butyl bromopropionate, 2-fluorobenzyl bromide and 2-(2-ethoxyethoxy)ethyl bromide. 